CN85101484B - Fixed cell carrier and process for its preparation - Google Patents
Fixed cell carrier and process for its preparation Download PDFInfo
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- CN85101484B CN85101484B CN 85101484 CN85101484A CN85101484B CN 85101484 B CN85101484 B CN 85101484B CN 85101484 CN85101484 CN 85101484 CN 85101484 A CN85101484 A CN 85101484A CN 85101484 B CN85101484 B CN 85101484B
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Abstract
The present invention relates to a new carrier for an immobilized cell, which is characterized in that the present invention comprises cells, sea tangle glue, calcium salt and water. Meanwhile, the present invention also discloses a method for manufacturing the new carrier for an immobilized cell and the application thereof.
Description
The present invention is a kind of new carrier and manufactures the method using.Particularly, in fixed cell technology, the present invention's carrier is the carrier for fixed cell, and it contains cell and water, it is characterized in that, it also contains seaweed gel and calcium carbonate.
Immobilized cell technology is the new technology, particularly immobilized yeast technology that the seventies develops rapidly, is a new biotechnology starting in recent years research.This technology can be widely used in medical science, biology, and microbiology, biotechnology, in the fields such as chemical engineering and foodstuffs industry.But at present, be still all in laboratory and pilot scale stage abroad.This be mainly in default of good carrier for many years all striving to find a kind of carrier, this carrier will meet following, that is: cheap, source is abundant, can produce in a large number, there is good solidity and toughness, nontoxic, can make the substrate of reaction and product be easy to come in and go out carrier and cell is difficult for passing through, cell speeds in carrier, growth is stable, and the life-span is long.But up to now, it is very not successful that people do various trials.Particularly cost is more expensive, cannot measure production, the solidity of carrier and poor toughness, and work-ing life is short, and yeast born of the same parents' propagation is not ideal enough.In order to address the above problem, the inventor a large amount of deep research, created a kind of new carrier, and now completed the present invention.Surprisingly, carrier of the present invention has many points, and it is cheap, and source is abundant, solidity and toughness
long with the life-span, nontoxic, yeast cell growth is good, increases
can come in and go out smoothly carrier and cell of thing and product is difficult for
carrier contain seaweed gel, calcium carbonate, cell and seaweed gel
as seaweed gel, but with the large sea-tangle of molecular weight for well, as point
be 150,000 left and right, the polymerization degree is 80 to 100 to be advisable, wherein better with the high person of D-MANNOSE aldehydic acid content, though routine produce in seaweed gel method, the intermediate before calcification.Carbonic acid can be ordinary calcium carbonate, but take more than chemical pure as better.Cell can select according to concrete object.Contained water, water that should common hygienic standard, as too much in foreign matter content in water, such as containing sulfurous gas etc., just should process in advance, to remove objectionable impurities.The content of sea glue is 1.5%～2.5% (weight percent), and 1.8% to 2.2% is (weight percent) well.Calcium carbonate containing being 0.5% to 2% (weight percent), with 0.8%1.2% (weight percent) for well.The content of cell, determine according to the kind specifically making, as select yeast cell, such as cereuisiae fermentum thin (Sccharomyces cereviside Hansen), Sucus Vitis viniferae ferment (Sccharomyccs uvarum Beijerinck), content answers 0.5% to 4%, and with 1.8% to 2.2% for well.The amount of water is 92.5% to 97.5%.In addition, if necessary, also can add the material such as fiber and chemical gel, when increasing the intensity of carrier and the tough carrier as the present invention as liquid state, it should show acid, and its pH value to be being about 3.5 to 6.5, and with 4 to 5 for well.The other feature of the present invention's body is that it is insoluble to polar solvent, as water etc.; It is insoluble to the solution containing ethanol, as the beer in drinks, fruit juice beer, gas, malt extract beverage, mlt extract syrup, light sparkling wine class.As little fragrant shore and grape wine, low alcohol white spirit, yellow rice wine and soy sauce, vinegar, it is also insoluble to organic acid containing in organic acid solution, as amino acid, xitix, lactic acid, first succinic acid and the solution that contains them; It is also insoluble to ketone, as the third butanone and containing ketone solution; Alcohols is as methyl alcohol, ethanol, propyl alcohol, sorbyl alcohol and alcoholic solution; In addition, it is also insoluble to syrup, sugary solution.Carrier of the present invention has good solidity and toughness.No matter transporting, in use procedure, when carrier is collided, when extruding, it can guarantee its integrity and unlikely fragmentation.
Carrier of the present invention also has another outstanding feature, i.e. its long service life general all can reach 70 to 100 days, and best reaches 120 days to 150 days.
Carrier of the present invention is nontoxic, uses carrier of the present invention and the product that produces, is can be because not used carrier of the present invention contaminated.Particularly to medicine, foodstuffs industry, it is to meet safety, hygienic requirements.
Another feature of the present invention is, completes after the immobilization of cell, and the propagation of cell in carrier is very fast, as immobilized yeast bacterium, and immobilized number of yeast cells, it is high more than 100 times that specific ionization is cultivated.
Moreover carrier of the present invention, in the time of immobilized yeast cell, also has such feature, in carrier, yeast is constantly bred, and the yeast that is attached to carrier surface is omitted in fermented liquid, and therefore, the yeast in carrier is always in newborn state.And Yeast proliferation amount is about 1000 times in carrier.
In addition, the present invention's raw material resources used are very abundant, and seaweed gel and calcium carbonate face upward and bow in a lot of countries and regions, thereby cost are very cheap, to the demand of the large production of industry, are can meet and without a doubt.
The cell that the present invention's carrier includes can be cereuisiae fermentum (Saccharomycos cereviside Hansen), saccharomyces uvarum (Saccharomycos uuvarum Berjerinck), acetone-butanol clostridium (Clostridium acetobutylicum), black acetobacter
The method of preparing the present invention's carrier comprises the following steps:
1. seaweed gel, calcium carbonate is soluble in water, and suitable solubility promoter in addition;
2. pair step 1 products therefrom carries out germicidal treatment;
3. pair step 2 products therefrom carries out cooling process;
4. in pair step 3 products therefrom, add cell and mix;
5. pair step 4 products therefrom carries out cooling process;
6. pair step 5 products therefrom carries out hardening treatment;
7. pair step 6 products therefrom carries out rinsing.
More particularly, seaweed gel used is with the large seaweed gel of molecular weight for well, and if molecular weight is 150,000, the polymerization degree is 80 to 100 seaweed gel.Calcium carbonate to be to have removed the calcium carbonate of impurity, or more than chemical pure calcium carbonate is for well.Solubility promoter with sodium-chlor for well, get seaweed gel 1.5% to 2.5%, take 1.8% to 2.2% as better, calcium carbonate 0.5% to 2%, take 0.8% to 1.2% as better, sodium-chlor 0.1% to 1%, take 0.2% to 0.4% as better, soluble in water, during as needs, stir so that they dissolve fully.
Gained solution is carried out to germicidal treatment.Be treated to heat-sterilization, Heating temperature is 65～85 ℃, and 75～80 ℃ better.Temperature is too low, does not reach the requirement of sterilization, and excess Temperature can cause bad consequence, and as sex change, coking etc., all can cause disadvantageous effect to the quality of the product of finally making.As use other sterilization processing instead, and although can reach the object of sterilization, having residual nuisance and be present in product, this is also very disadvantageous.Remove these residues and can expend more multi-step and equipment, therefore, be not inclined to these methods of using.If residue has toxicity, more can affect normal growth and the breeding of cell, remove this this Toxic, will increase expenditure, thereby more should avoid as possible.
To carry out cooling process through the product of heat-sterilization processing, the method for cooling of available routine, as naturally cooling, no matter ventilate cooling and circulating water etc., adopt any method, finally will make solution temperature be down to 15～35 ℃, take 25～30 ℃ as better.Because in this temperature range, heating will be fixing cell, can make cell itself be without prejudice, too low, too high temperature, all can on to cause totally unfavorable impact by fixing cell.If cell used itself has special requirement, certainly within the rule.If cell used is yeast cell, consumption is 0.5%～4%, and with 1.8%～2.2% for well, this amount ranges, the concentration that is yeast suspension is 10/ milliliter to be calculated and uses consumption, and in fact used be exactly the suspension of yeast cell.For different cells, carry out depending on particular case.If the quantity of yeast cell is too little, unfavorable to completing production; If yeast cell consumption is excessive, can not guarantees their growths, the synchronism of growing, breeding, thereby make the functional lability of fixed cell carrier, thereby cause unstable product quality.Add after cell, as needs mix, the method for available routine completes.
The carrier soln that is mixed with cell is put into former, by the mode of injection or water clock, make this carrier soln enter calcium chloride solution, form water-fast gel particle.The diameter of the particle forming is 1.5～4mm, take 2～3mm as better.The strength of solution of calcium chloride used is 2%～4%, with 2.5%～3% for well.
By the carrier forming, put into calcium chloride solution and soak, the concentration of calcium chloride solution used is 2%～4%, with 2.5%～3% for well.15～35 ℃ of temperature requirements, with 25～30 ℃ for well.The time of hardening treatment is more than 30 minutes, with 2～8 hours for well.Through the carrier of hardening treatment, its solidity and toughness are all good.Such fixed yeast particle, from calcium chloride solution, take out, by rinsed with sterile water, until muriate is rinsed totally.
Like this, just make the present invention's carrier.Made fixed yeast particle can directly apply to production, also can be placed in suitable fermented liquid, cultivate for some time for the production of.
Because the present invention's carrier has advantages of uniquely, thereby the scope of its use is quite extensive.It may be used on, in medicine production, being particularly used in and utilizing microorganism cells to produce, or transform out expensive microbiotic or other medicines.
It also can be applicable in microbiology field, as produces a large amount of microorganisms, and the microorganism of new generation is cultivated rapidly.
Particularly, in biotechnology, due to the present invention's carrier, make the new microorganism obtaining by biotechnology, go among can being applied to actual production, thereby benefit society.
Because the present invention's carrier is nontoxic, so it is particularly suitable for being applied in foodstuffs industry, as produce each seed amino acid, vitamins c etc.
As one of topmost Application Areas of the present invention, it is brewery industry.The present invention's carrier, is yeast as what comprise, is just specially adapted to manufacture various drinks, as Fermented Drinks such as beer, beer-like beverage, fruit juice beer, Ge Wasi, mlt extract syrup, malt extract beverage, light sparkling wine, little champagne.In addition, be also applicable to manufacture the fermented soy industry such as yellow rice wine, low alcohol white spirit, soy sauce, vinegar.
In the production process of making beer, wherein a crucial step is Primary Fermentation (ferment before claiming again).Traditional technology in the past, all ferment at fermentation vat (or in tank) with free culturing yeast bacterium, the time of fermentation is generally 7～12 days, utilize the present invention's fixed yeast, can make time shorten to 20～36 hour of Primary Fermentation, and can use continuously 70～100 days, best situation can be used 20～150 days.When fermentation, temperature is controlled at 6～9 ℃.In addition, while using the present invention's Immobilized Yeast beer, also make the rear ferment time within 30～60 days, shorten to 8～15 days by traditional technology.The cycle of beer production is shortened, and plant factor improves, and brings very large economic benefit.
What is more important, due to the birth of the present invention's fixed yeast, makes the serialization of beer production, and automatization becomes possibility.Substrate is sent into and reverse rising by the bottom of the fermentation container that fixed yeast is housed, during this period, by fixed yeast, substrate is reacted, in the time that product is qualified, emitted by the top of reactor, control the inlet of substrate and the discharge of product simultaneously, be as the criterion to reach optimum regime.The production serialization of beer is just achieved like this.If use corresponding machinery to carry out the steps such as pan feeding, discharging so just to have completed the continuous and automatic of beer production simultaneously.Will carry out a revolution to existing process for beer production like this, can save a large amount of funds simultaneously, equipment, manpower, improves plant factor, well imagines, and it is huge that the benefit that uses the present invention's fixed yeast to bring is bound to.
In addition, when traditional technology is produced yellow rice wine in the past, the front ferment time was generally about three months, and used the present invention's fixed yeast, and the front ferment time is general only with 60 hours.After traditional technology, the ferment time is 90 days, and adopts the present invention's fixed yeast, and the rear ferment time is only one month.And owing to using the present invention's fixed yeast, make the production of yellow rice wine except original batch-type, also can adopt continous way.Like this, apparent, use the present invention's fixed yeast, the income of bringing in yellow rice wine process industry, is considerable.
In the past, because fixed yeast can not be applied in industrial production, particularly in fermentation industry, so that the production of white wine is confined to produce the white wine of the high number of degrees.Because when producing when white wine by traditional technology, the number of degrees of the wine of production are too low, will make the wine of output with great unpalatable shortcoming, local flavor is also bad, taste also not alcohol and.These are all because wherein containing having a large amount of potato spirit, aldehydes, ester class to cause.And traditional technology cannot solve the problem of this respect.If but use the present invention's fixed yeast just can overcome above-mentioned technical difficulty.Produce in the process of white wine in traditional technology, ferment with the present invention's fixed yeast, the low alcohol white spirit that just can make institute's output is less containing potato spirit, aldehydes, ester class, the taste pure of wine with, alcoholic strength is spent 25～50.The low alcohol white spirit of suitable especially manufacture 25～38 degree.Simultaneously, owing to using fixed yeast, make the production of low alcohol white spirit also can become serialization, automatization, as can be seen here, use the present invention's fixed yeast, producing containing potato spirit, aldehydes, when the less low alcohol white spirit of ester class, the technique that yet makes white wine produce has obtained huge improvement, and income is difficult to the appraisal.
In addition, the present invention also can be used in production vinous.Production vinous in the past, is mainly band Pericarpium Vitis viniferae fermentation, is mainly that to utilize the wild yeast of Pericarpium Vitis viniferae be starter.The grape wine that produced like this, because the tannin anthocyanogen in Pericarpium Vitis viniferae is all dissolved in Sucus Vitis viniferae, extends the rear ferment phase.Particularly, in the time producing white wine, cause the difficulty keeping in transparency, in addition, use wild yeast to be also difficult to guarantee safety in production.If adopt the present invention's fixed growth yeast, can make fermenting speed accelerate, the front ferment phase can be down to by original 7 days about 24 hours, and the rear ferment phase can be down to about 3 months by original half a year.Meanwhile, also give safety in production vinous with strong assurance.Particularly producing when white wine, can be without original wild yeast in Pericarpium Vitis viniferae as starter, thereby just can remove Pericarpium Vitis viniferae, the color of white wine is more easily guaranteed.
Moreover the present invention's fixed yeast, in the time of the production technique of the traditional sauce oil and vinegar of transformation, thereby has shortened the time of fermenting greatly because it has substituted fermentation step in the past, has also shortened widely the production cycle.Compared with traditional technology, can raise the efficiency tens times.And the product of producing, the product that can go out with traditional explained hereafter completely compares favourably.
Following examples further illustrate the present invention, in any case but also do not limit the present invention.
Embodiment 1
By seaweed gel, calcium carbonate, sodium-chlor (by above-mentioned weight) is added in water (94.6 kilograms) and stirring and evenly mixing, with 75 ℃ of heat-sterilizations, is then cooled to room temperature, by 2 liter (10
8individual/milliliter) yeast cell suspension adds wherein, after stirring and evenly mixing, put into former, splash in 3% calcium chloride solution and make it into pellet shapes, after shaping, it is that the calcium chloride solution that the concentration of 28 ℃ is 2.5% soaks 4 hours that carrier granule is put into temperature.Then take out carrier granule rinsed with sterile water, obtained the cerevisiae that contains of the present invention, the carrier of calcium carbonate and seaweed gel, this carrier has good solidity and toughness.
Embodiment 2
Be prepared into cell carrier of the present invention with above-mentioned each component by the method described in embodiment 1, the solidity of this carrier and toughness are all fine.
Embodiment 3
Make carrier granule of the present invention by the method described in embodiment 1 by said components, the solidity of carrier and toughness are all fine.
Embodiment 4
Press the method described in embodiment 1, get seaweed gel 2%, calcium carbonate 2%, cereuisiae fermentum suspension (10
8individual/milliliter) 2%, sodium-chlor 0.3%, water 93% is made 10 cubic metres of carrier granule total amounts of the present invention.Carrier granule is all packed in the fermentor tank of 60 measurement tons, in tank, be divided into five layers with sieve plate, every layer packs 2 tons of fixed growth yeast particles (amounting to 5 × 2 tons=10 tons) prepared by carrier of the present invention into.Wort is entered from fermentor tank bottom, and flow path granular layer is discharged to the outlet at fermentor tank top from the bottom to top, and flow is 0.4 liter/second, flows out 34 tons of bright beers every day.Use 100 days continuously from August 1 nineteen eighty-three to November 11 then, while implementing to finish, carrier granule is still intact, thus need not be as routine techniques in the past frequent replacing yeast, thereby the serialization of beer production energy is carried out over a long time.This enforcement is carried out in brewery of Lingbi county of Anhui Province.
Embodiment 5
Get seaweed gel (2%), calcium carbonate (2%), cereuisiae fermentum (Denmark's Carlsberg yeast, 2%), sodium-chlor (0.9%), water (93.1%) is by being prepared into 10 cubic metres of carrier granules of the present invention described in embodiment 1.Pack large ferment tank into by the method described in embodiment 5 and manufacture beer.Move and amount to 116 days continuously from February 27,3 days to 1985 November in 1984.When enforcement stops, carrier granule is still intact.In Beijing, (Shunyi, Beijing) brew-house carries out in this enforcement.
Embodiment 6
Make 10 tons of fixed yeast particles with above-mentioned formula by the method described in embodiment 1, carry out large-scale experiment, approximately 3200 tons of common property beer, reach 100 days the work-ing life of carrier.Product meets the requirements.
In process of the test, to fermentation time and temperature, pol, pH value, the variation of yeast amount etc. is measured, and the results are shown in Table 1.
The presentation of results of table 1, in fermenting process, should keep pH between 4 to 5.5 for well.In addition, after reaction proceeds to 24 hours, product basically forms, and limit fermentation degree is 78%, the minimum pol 2.6 of fermenting.And temperature should be controlled at 6～10 ℃ for well.
Generated beer is tested, and acquired results is in table 2:
Table 2
The presentation of results of table 2, the beer that immobilized yeast technology is produced meets the requirement of beer production completely.
In addition, owing to using the present invention's immobilized yeast, not only obtain the improvement of producing beer technology, produce qualified beer, there is another wonderful effect simultaneously, no matter the amino acid whose content of the beer produced, higher than the beer of traditional technology production, be to produce beer or pasteurized beer is not always the case.Concrete measurement result is in table 3, table 4:
Can be learnt clearly by table 3, use the aminoacids content of the unpasteurized beer of immobilized yeast technology manufacturing, the content of the beer of producing than traditional technology is a lot.
Can be learnt by table 4, except tryptophane, outside Serine and Threonine, remaining amino acid is all take the aminoacids content of Immobilized Yeast as high.
Reference embodiment
Get sodium alginate 4% (250 kilograms), yeast cell suspension (10
8individual/milliliter) 2%, with appropriate water routinely technique make the carrier granule of embedding yeast.To manufacturing beer with made particle fermentation July 24 then, amount to 48 days from June 6 nineteen eighty-two.When experiment finishes, carrier granule breaks, cannot continue to use.
Table 1
Table 3
Table 4
Claims (9)
1. a fixed cell carrier, it contains cell and water, it is characterized in that, and it also contains the seaweed gel of 1.5-2.5% (weight) and the calcium carbonate of 0.5-2% (weight).
2. carrier according to claim 1, wherein said seaweed gel is the seaweed gel of molecular weight in 150,000 left and right.
3. according to the carrier described in claim 1, wherein said cell is yeast cell, preferably beer yeast cell (Saccharomyces cereviside) and saccharomyces uvarum (Saccharomyces uvarum) cell.
4. according to the carrier described in any one in claim 1 or 3, wherein said cell content is that 0.5-4% (weight) is (with every ml yeast suspension containing 10
8individual cell calculates).
5. the method for preparation carrier as described in claim 1, is characterized in that, the method comprises the following steps:
1), by seaweed gel, calcium carbonate and sodium-chlor are added to the water and mix;
2) to step 1) product carry out germicidal treatment;
3) to step 2) product carry out cooling process;
4) in step 3) product in add cell;
5) to step 4) product form processing;
6) to step 5) product carry out hardening treatment;
7) to step 6) product float Xian.
6. according to the method described in claim 5, the wherein said consumption that adds seaweed gel is 1.5-2.5% (weight).
7. according to the method described in claim 5, the wherein said consumption that adds calcium carbonate is 0.5-2% (weight).
8. according to the method described in claim 5, the wherein said cell preferred yeast cell adding.
9. the application of carrier in fermentation industry especially brewing industry as described in claim 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 85101484 CN85101484B (en) | 1985-04-01 | 1985-04-01 | Fixed cell carrier and process for its preparation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 85101484 CN85101484B (en) | 1985-04-01 | 1985-04-01 | Fixed cell carrier and process for its preparation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN85101484A CN85101484A (en) | 1986-07-30 |
| CN85101484B true CN85101484B (en) | 1987-01-14 |
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ID=4791864
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 85101484 Expired CN85101484B (en) | 1985-04-01 | 1985-04-01 | Fixed cell carrier and process for its preparation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN85101484B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101544944B (en) * | 2008-03-29 | 2013-01-23 | 杨迪 | Novel method for accelerating aging and impurity removal of white spirit by immobilized enzyme |
| JP5414090B2 (en) * | 2008-10-09 | 2014-02-12 | 独立行政法人農業・食品産業技術総合研究機構 | Solid support on which microorganism group performing parallel dual mineralization reaction is immobilized, catalyst column, and method for producing solid medium for plant cultivation |
-
1985
- 1985-04-01 CN CN 85101484 patent/CN85101484B/en not_active Expired
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| Publication number | Publication date |
|---|---|
| CN85101484A (en) | 1986-07-30 |
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